Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 108, Issue 2, Pages 395-403Publisher
WILEY
DOI: 10.1002/bit.22916
Keywords
cellular array; transcription factor activity; bioluminescence imaging
Categories
Funding
- NIH [R21 006520, T32 GM008449]
- Chicago Biomedical Consortium
- The Chicago Community Trust
Ask authors/readers for more resources
Identifying molecular mechanisms or therapeutic targets is typically based on large-scale cellular analysis that measures the abundance of mRNA or protein; however, abundance does not necessarily correlate with activity. We report a method for direct large-scale quantification of active pathways that employs a cellular array with parallel gene delivery of constructs that report pathway activity. The reporter constructs encode luciferase, whose expression is influenced by binding of transcription factors (TFs), which are the downstream targets of signaling pathways. Luciferase levels are quantified by bioluminescence imaging (BLI), which allows for rapid, non-invasive measurements. Activity profiles by BLI of 32 TFs were robust, consistent, and reproducible, and correlated with standard cell lysis techniques. The array identified five TFs with differential activity during phorbol-12-myristate-13-acetate (PMA)-induced differentiation of breast cancer cells. A system for rapid, large-scale, BLI quantification of pathway activity provides an enabling technology for mechanistic studies of cellular responses and processes. Biotechnol. Bioeng. 2011;108: 395-403. (C) 2010 Wiley Periodicals, Inc.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available